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Diagnosis and Direct Automated Sequencing of HIV-1 From Dried Blood Spots (DBS) Collected on Filter Paper

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HIV Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 17))

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Abstract

Since its discovery in 1981, human immunodeficiency virus type 1 (HIV-1) has rapidly emerged as one of the most devastating infectious pathogens of this century (13). The World Health Organization (WHO) estimates that, as of 1995, there were at least 15 million HIV- infected men, women, and children worldwide, with the vast majority of infections occurring in developing countries and isolated rural regions where specimen collection, preparation and shipment are difficult (4). Simple and improved sampling methods that can be widely applied under difficult field conditions are needed to effectively monitor the changing dynamics of the HIV-1/AIDS pandemic, track the spread of HIV-1 variants among different population groups, and ensure that research and interventive activities are directed against biologically important variants of the virus. To date, at least eight major HIV-1 subtypes, designated A through H, have been identified (5,6). More recently, a ninth subtype, I, has been detected (7), as well as several highly divergent, or “outlying” variants of HIV-1 that have been tentatively classified as group O (8,9). This subtyping is based on a relatively small number of specimens collected from a few geographic areas and the full range and distribution of HIV-1 variants remains to be established. The collection of whole blood on filter paper provides an innovative and powerful approach for the systematic and unbiased collection of large numbers of field specimens for diagnostic and surveillance purposes (1019).

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References

  1. Lederberg, J., Shope, R. E., and Oaks, S., Jr., (eds.) (1992) Emerging Infections: Microbial Threats to Health in the United States. National Acadermy of Sciences, Washington, DC.

    Google Scholar 

  2. Hu, D. J., Dondero, T. J., Rayfield, M. S., George, J. R., Schochetman, G., Jaffe, H. W., et al. (1996) The emerging genetic diversity of HIV. The importance of global surveillance for diagnostics, research and prevention. JAMA 275, 210–215.

    Article  PubMed  CAS  Google Scholar 

  3. Weniger, B. B., Takebe, Y., Ou, C-Y., and Yamazaki, S. (1994) The molecular epidemiology of HIV in Asia. AIDS 8(Suppl 2), S13–S28.

    PubMed  Google Scholar 

  4. Nowak, R. (1995) Staging ethical AIDS trials in Africa. Science 269, 1332–1335.

    Article  CAS  Google Scholar 

  5. Myers, G., Korber, B., Wain-Hobson, S., Smith, R. F., and Pavlakis, G. N. (1993) Human Retroviruses and AIDS. Los Alamos National Laboratory, Los Alamos, New Mexico.

    Google Scholar 

  6. Louwagie, J., McCutchan, F. E., Peeters, M., Brennen, T. P., Buell, E., Eddy, G. A., et al. (1993) Phylogenetic analysis of gag genes from 70 international HIV-1 isolates provides evidence for multiple genotypes. AIDS 7, 769–780.

    Article  PubMed  CAS  Google Scholar 

  7. Kostrikis, L. G., Bagdades, E, Cao, Y, Zhang, L., Dimitriou, D., and Ho, D. D. (1995) Genetic analysis of human immunodeficiency virus type 1 strains from patients in Cyprus: identification of a new subtype designated subtype I. J Virol. 69, 6122–6130.

    PubMed  CAS  Google Scholar 

  8. Gurtler, L. G., Hauser, P. H., Eberle, J., von Brunn, A., Knapp, S., Zekeng, L., Tsague, J. M., and Kaptue, L. (1994) A new subtype of human immunodeficiency virus type 1 (MVP-5180) from Cameroon. J. Virol. 68, 1581–1585.

    PubMed  CAS  Google Scholar 

  9. DeLeys, R., Vanderborght, B, Vanden Haesevelde, M., van Geel, A., Wauters, C., et al. (1990) Isolation and partial characterization of an unusual human immunodeficiency retrovirus from two persons of West-Central African origin. J. Virol. 64, 1207–1216.

    CAS  Google Scholar 

  10. Garrick, M. D., Dembure, B. S., Cuthrie, R. (1973) Sickle-cell anemia and other hemoglobinopathies: procedures and strategy for screening employing spots of blood on filter paper as specimens. N. Engl. J. Med. 288, 1256–1268.

    Article  Google Scholar 

  11. McCabe, E. R. B., Huang, S.-Z., Seltzer, W. K., and Law, M. L. (1987) DNA microextraction from dried blood spots on filter paper blotters: potential applications to newborn screening. Hum. Genet. 75, 213–216.

    Article  PubMed  CAS  Google Scholar 

  12. Hoff, R., Berarde, V. P., Weiblen, B. J., Mahoney-Trout, L, Mitchell, M., and Grad, G. (1988) Seroprevalence of human immunodeficiency virus among child-bearing women: estimation by testing samples of blood from newborns in New York State. N. Engl. J. Med. 318, 525–530.

    Article  PubMed  CAS  Google Scholar 

  13. Cassol, S., Salas, T., Arella, M., Neumann, P., Schechter, M., and O’Shaughnessy, M. (1991) Use of dried blood spot specimens in the detection of human immunodeficiency virus type 1 by the polymerase chain reaction. J. Clin. Microbiol. 29, 667–671.

    PubMed  CAS  Google Scholar 

  14. Cassol, S., Salas, T., Gill, M. J., Montpetit, M., Rudnik, J., Sy, C. T., et al. (1992) Stability of dried blood spot specimens for detection of human immunodeficiency virus DNA by polymerase chain reaction. J. Clin. Microbiol. 30, 3039–3042.

    PubMed  CAS  Google Scholar 

  15. Cassol, S. A., Lapointe, N., Salas, T., Hankins, C., Arella, M., Fauvel, M., et al. (1992) Diagnosis of vertical HIV-1 transmission using the polymerase chain reaction and dried blood spot specimens. J. Acq. Immun. Defic. Syndr. 5, 113–119.

    CAS  Google Scholar 

  16. Cassol, S., Butcher, A., Kinard, S., Spadoro, J., Sy, T., Lapointe, N., et al. (1994) Rapid screening for the early detection of mother-to-child transmission of HIV-1. J. Clin. Microbiol. 32, 2641–2545.

    PubMed  CAS  Google Scholar 

  17. Cassol, S., Weniger, B. G., Babu, P. G. Salminen, M., Zheng, X., Htoon, M. T., et al. (1996) Detection of HIV-1 env subtypes A, B, C and E in Asia using dried blood spots: a new surveillance tool for molecular epidemiology. AIDS Res. Hum. Retrovir. 12, 1435–1441.

    Article  PubMed  CAS  Google Scholar 

  18. Cassol, S., Read, S., Weniger, B G., Gomez, P., Lapointe, N., Ou, C-Y., et al. (1996) Dried blood spots collected on filter paper: an international resource for the diagnosis and genetic characterization of HIV-1. Mem. Inst. Oswaldo Cruz, Rio de Janeiro, 91, 351–358.

    CAS  Google Scholar 

  19. Cassol, S. (1996) Applications of dried blood spots to the diagnosis and field surveillance of HIV-1. Benchmark 3, 7–9.

    Google Scholar 

  20. Knudsen, R. C., Slazyk, W. E., Richmond, J. Y., and Hannon, W. H. (1993) Guide-lines from the Centers for Disease Control and Prevention (Atlanta) for the shipment of dried blood spot specimens. Infant Screening 16, 1–3. FAX Information Service (404-332-4565). Document no. 101011, 13 October.

    Google Scholar 

  21. Folks, T. M., Powell, D., Lightfoote, M., Koenig, S., Fauci, A. S., Benn, S., et al. (1986) Biological and biochemical characterization of a cloned Leu-3-cell surviving infection with the acquired immune deficiency syndrome retrovirus. J. Exp. Med. 164, 280–290.

    Article  PubMed  CAS  Google Scholar 

  22. National Committee for Clinical Laboratory Standards. NCCL Approved Standard LA4-A2. (1992) Blood collection on filter paper for neonatal screening programs. National Committee for Laboratory Standards, Villanova, PA.

    Google Scholar 

  23. Harvey, M. A., King, T. H., and Burghoff, R. (1996) Impregnated 903 blood collection paper. A tool for DNA preparation from dried blood spots for PCR amplification. A technical Bulletin from the Research and Development, Schleicher & Schuell, Inc., Keene, NH 03431; Fax (603) 357-3627.

    Google Scholar 

  24. Singer-Sam, J., Tanguay, R. L., and Riggs, A. D. (1989) Use of Chelex to improve the PCR signal from a small number of cells. Amplifications 3, 11.

    Google Scholar 

  25. Longo, M. C., Beringer, M. S., and Hartley, J. L. (1990) Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene 93, 125–128.

    Article  PubMed  CAS  Google Scholar 

  26. Kwok, S. (1990) Procedures to minimize PCR-product carry-over, in PCR Protocols: A Guide to Methods and Application, (Innis, M. A., Gelfand, S. H., Sninsky, J. J., and White, T. J. eds), Academic Press, San Diego, CA, pp. 142–145.

    Google Scholar 

  27. Perkin Elmer Corporation, Applied Biosystems Division. (1995) Comparative PCR Sequencing: A Guide to Sequencing-Based Mutation Detection. Foster City, CA.

    Google Scholar 

  28. Smith, S., Overbeek, R., Woese, C. R. Gilbert, W., and Gillevet, P. (1994) The genetic Data environment: an expandable GUI for multiple sequence analysis. Comp. Appl. Biol. Sci. 10, 671–675.

    CAS  Google Scholar 

  29. Olsen, G. J., Matsude, H., Hagstrom, R., and Overbeek, R. (1994) FastDNAml: A tool for construction of phylogenetic trees of DNA sequences using maximum likelihood. Comput. Appl. Biosci. 10, 41–48.

    PubMed  CAS  Google Scholar 

  30. Cassol, S., Gill, M. J., Pilon, R., Cormier, M., Voight, R. F., Willoughby, B., and Forbes, J. (1997) Quantification of human immunodeficiency virus type 1 RNA frim dried plasma spots collected on filter paper. J. Clin. Microbiol. 35, 2795–2801.

    PubMed  CAS  Google Scholar 

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© 1999 Humana Press Inc., Totowa, NJ

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Cassol, S.A., Read, S., Weniger, B.G., Pilon, R., Leung, B., Mo, T. (1999). Diagnosis and Direct Automated Sequencing of HIV-1 From Dried Blood Spots (DBS) Collected on Filter Paper. In: Michael, N.L., Kim, J.H. (eds) HIV Protocols. Methods in Molecular Medicine™, vol 17. Humana Press. https://doi.org/10.1385/0-89603-369-4:125

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  • DOI: https://doi.org/10.1385/0-89603-369-4:125

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-369-6

  • Online ISBN: 978-1-59259-601-0

  • eBook Packages: Springer Protocols

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